Sprouty2 inhibition promotes proliferation and migration of periodontal ligament cells

Urara Tanaka, Terukazu Sanui, Takao Fukuda, Kyousuke Toyoda, T. Taketomi, R. Atomura, K. Yamamichi, Hidefumi Maeda, Fusanori Nishimura

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Objectives: We previously demonstrated that a dominant-negative Sprouty2 (Spry2) mutation promotes osteoblast proliferation and differentiation after basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) stimulation, whereas it diminishes proliferation of gingival epithelial cells, thereby inducing favourable conditions for periodontal tissue regeneration. In this study, we investigated how Spry2 inhibition affects the cellular physiology of periodontal ligament (PDL) cells. Methods: A total of 1-17 PDL cells (multipotent clonal human PDL cell line) were stimulated with bFGF and EGF after transfection of Spry2 siRNA. Cell proliferation, migration, ALP staining, real-time PCR, Western blot and immunofluorescence assays were performed. Results: ERK1/2 activation and proliferation of 1-17 PDL cells were significantly upregulated by the addition of Spry2 siRNA in the presence of bFGF and EGF. In addition, Spry2 siRNA reduced transcription of osteogenesis-related genes and ALP staining relative to control cells. Furthermore, it increased AKT/phosphatidylinositol 3-kinase (PI3K) phosphorylation; consequently, Rac1 but not Cdc42 was activated, thereby promoting lamellipodia formation, cell proliferation and migration after stimulation by bFGF and EGF. Conclusion: Spry2 combined with bFGF and EGF stimulation reduced PDL cell migration and proliferation with inducing osteoblastic differentiation. These in vitro findings may provide a molecular basis for novel therapeutic approaches for establishing periodontal tissue regeneration.

Original languageEnglish
Pages (from-to)977-986
Number of pages10
JournalOral Diseases
Volume21
Issue number8
DOIs
Publication statusPublished - Nov 1 2015

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Periodontal Ligament
Fibroblast Growth Factor 2
Epidermal Growth Factor
Small Interfering RNA
Cell Movement
Cell Proliferation
Regeneration
Phosphatidylinositol 3-Kinase
Staining and Labeling
Pseudopodia
Osteoblasts
Osteogenesis
Fluorescent Antibody Technique
Transfection
Real-Time Polymerase Chain Reaction
Western Blotting
Epithelial Cells
Phosphorylation
Cell Line
Mutation

All Science Journal Classification (ASJC) codes

  • Otorhinolaryngology
  • Dentistry(all)

Cite this

Sprouty2 inhibition promotes proliferation and migration of periodontal ligament cells. / Tanaka, Urara; Sanui, Terukazu; Fukuda, Takao; Toyoda, Kyousuke; Taketomi, T.; Atomura, R.; Yamamichi, K.; Maeda, Hidefumi; Nishimura, Fusanori.

In: Oral Diseases, Vol. 21, No. 8, 01.11.2015, p. 977-986.

Research output: Contribution to journalArticle

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abstract = "Objectives: We previously demonstrated that a dominant-negative Sprouty2 (Spry2) mutation promotes osteoblast proliferation and differentiation after basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) stimulation, whereas it diminishes proliferation of gingival epithelial cells, thereby inducing favourable conditions for periodontal tissue regeneration. In this study, we investigated how Spry2 inhibition affects the cellular physiology of periodontal ligament (PDL) cells. Methods: A total of 1-17 PDL cells (multipotent clonal human PDL cell line) were stimulated with bFGF and EGF after transfection of Spry2 siRNA. Cell proliferation, migration, ALP staining, real-time PCR, Western blot and immunofluorescence assays were performed. Results: ERK1/2 activation and proliferation of 1-17 PDL cells were significantly upregulated by the addition of Spry2 siRNA in the presence of bFGF and EGF. In addition, Spry2 siRNA reduced transcription of osteogenesis-related genes and ALP staining relative to control cells. Furthermore, it increased AKT/phosphatidylinositol 3-kinase (PI3K) phosphorylation; consequently, Rac1 but not Cdc42 was activated, thereby promoting lamellipodia formation, cell proliferation and migration after stimulation by bFGF and EGF. Conclusion: Spry2 combined with bFGF and EGF stimulation reduced PDL cell migration and proliferation with inducing osteoblastic differentiation. These in vitro findings may provide a molecular basis for novel therapeutic approaches for establishing periodontal tissue regeneration.",
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AU - Fukuda, Takao

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AU - Yamamichi, K.

AU - Maeda, Hidefumi

AU - Nishimura, Fusanori

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